Listen In - Bitesize Bio Webinar Audios: Recent Episodes
Bitesize Bio
The Listen In podcast from Bitesize Bio is a compilation of our best webinars to enjoy at your leisure, wherever and whenever.
Each episode is an opportunity to gain the valuable insights you need to advance your research.
From a crash course in developing fool-proof ELISAs to the latest applications and innovations in CRISPR/Cas9 and microscopy techniques, and much more—you'll hear about challenges encountered and discover practical solutions to achieve the best possible results.
Tap into the experience and expertise of leading researchers and commercial specialists to drive your research projects forward efficiently and productively. Listen In now!
https://bitesizebio.com/listen-in/
What's more disappointing than running your flow cytometry sample to discover it's rubbish? Especially considering time on cytometers can be precious, even more so if you've booked onto an in-demand one at a busy core facility.
You've done all the work and all the waiting—for nothing.
If this scenario sounds familiar, this episode of Listen In is for you.
Jessica Rowley, Flow Cytometry Facility Manager at Imperial College London, explains how to prepare flow samples optimized for your experiments, to save you time and avoid disappointment.
She has worked with hundreds of samples, so she is the perfect person to tell you how to optimize your flow cytometry protocol, from creating viable single-cell suspensions to staining, fixation, and improving data quality.
Plus, if you're new to flow cytometry, designing a new experiment, or want to improve your data quality, she has some advice on those, too.
Watch the full presentation here: https://events.bitesizebio.com/garbage-in-garbage-out-sample-prep
Browse all episodes of our educational webinar series here: https://listen-in.bitesizebio.com/
79 — Scientific writing is an art that requires as much attention and proficiency as your hands-on lab work. When done right, it can significantly contribute to your scientific endeavors. But, if not taken seriously, inconsistent language, formatting, and terminology can hinder your work's clarity and distract from your remarkable scientific findings.
In this episode of Listen In, our Content Creation Manager, Laura Grassie, unravels the complexities of consistent scientific writing and helps you steer clear of common pitfalls that make your work look sloppy. She introduces you to powerful, simple strategies that can significantly enhance the coherence, clarity, and, most importantly, the consistency of your research communication.
This episode is particularly beneficial if you're preparing for a crucial grant submission, planning your next research paper, or setting out to work on your thesis. The insights Laura shares will arm you with effective tools and techniques to achieve error-free, consistent scientific writing.
Don't let unorganized writing overshadow your extraordinary scientific work. Dive into this enlightening episode to discover the secret to consistent scientific writing that impresses everyone, from viva examiners to grant and paper reviewers.
Watch the full presentation here: https://events.bitesizebio.com/consistent-error-free-writing-tips/join
Browse all episodes of our educational webinar series here: https://listen-in.bitesizebio.com/
Are you an early career scientist looking for guidance on your career path? Or you're a seasoned researcher considering a career change and want to know what's out there.
We've brought together a panel of experts and bioscientists with a wide range of experience to share their insights and help you carve out the best career possible.
Get advice and tools to forge a path that suits you—inside or outside academia.
Our panel shares their personal stories and insights into the various career paths available to bioscientists. Plus, gain valuable insights and a new perspective on the rewarding possibilities for your future.
With:
• Professor Stuart Maudsley, Odysseus Professor of Receptor Pharmacology, University of Antwerp
• Dr. Jane Luff, Director of Delivery, Our Future Health UK
• Dr. Axel Thomson, Business Development Executive, Edinburgh Innovations
Watch the full presentation here: https://events.bitesizebio.com/lab-meetings-live-career-paths-for/room
Browse all episodes of the Listen In Series here: https://listen-in.bitesizebio.com/
Freeze fracture is a powerful technique that enables imaging of internal structures of cells and tissues at the nanoscale level by freezing samples at ultra-low temperatures and then fracturing them along natural weak points. These structures can be imaged without dehydration or distortion using cryo-SEM, providing high-resolution images with great clarity.
Discover how to use freeze fracture and cryo-SEM workflow to answer research questions and learn its exciting applications in cell biology.
Watch the full presentation here: https://microscopyfocus.com/breaking-the-ice-freeze-fracture/
Browse all episodes of the Listen In Series here: https://listen-in.bitesizebio.com/
As a researcher, one of the most crucial skills to develop is the ability to perform a thorough review of manuscripts. Manuscript reviewing is a vital job that requires specific skills and knowledge to properly assess published literature. By reading and reviewing papers, researchers can develop and hone these skills, which will help them identify and handle tricky issues such as image and data manipulation.
In this episode of the Listen In series, Dr. Johanna Ahlskog discusses the essential skills all reviewers and researchers need. She emphasizes the critical importance of the review process and offers general considerations when reviewing articles. Dr. Ahlskog also provides information on how to properly assess presented data and discusses issues surrounding image manipulation, including permissible versus problematic manipulation.
If you're interested in enhancing your manuscript reviewing skills, this episode is a must-listen. You'll gain valuable insights into the review process and learn how to navigate complex issues that may arise during the evaluation of a manuscript.
Watch the full presentation here: https://www.youtube.com/watch?v=8Nc5361cuTM
Browse all episodes of the Listen In Series here: https://listen-in.bitesizebio.com/
Have you ever wondered what other jobs are ideal matches for your analytic and artistic skills? Are you looking for an exciting new career in marketing, or have you ever wondered if it was for you?
In this episode of Listen In, discover how the assets that make you a great scientist could also make you a great marketer in today's data-driven markets.
Learn the role of a marketer, how scientific skills translate into this career in today's data-driven markets, and how to adapt your talents to achieve success!
Join us, and explore the world of marketing and the dynamic and dominant role it plays in society.
To view the full presentation of this webinar, click here: https://www.youtube.com/watch?v=an0v67ZzxbQ
Browse all episodes of the Listen In Series here: https://bitesizebio.com/listen-in/
Although macrophages were first described by Elie Metchnikoff in 1882, plenty of mysteries are still associated with the cell type. Indeed, while macrophages were once considered simply ""garbage trucks"" of the immune system due to their phagocytic property, their substantial and multifaceted contribution to immunological responses and homeostasis is becoming more apparent. Macrophages can produce a wide range of cytokines and chemokines to influence the immune response toward healing or inflammation. In as such, they possess a great deal of plasticity to respond with either pro- or anti-inflammatory signals depending on the environmental milieu. Moreover, researchers are beginning to turn to macrophages to assist chimeric antigen receptor (CAR) T cells in various immunotherapies.
This webinar is presented by Anne Lodge, Chief Scientific Officer of Astarte Biologics.
Functional imaging is a rapidly growing field key to driving new understanding in biology. Insights into the function and interaction of molecules are the key to reveal the underlying cellular mechanisms. In this context, fluorescence lifetime imaging (FLIM) is a powerful tool, providing valuable information beyond spectral imaging. FLIM is immune to concentration artefacts and sensitive to molecular environment such has pH changes, ion concentrations, and more.
Förster Resonance Energy Transfer (FRET) is an example of molecular environmental changes. The donor lifetime is shortened by the presence of the acceptor. FRET experiments thus benefit from FLIM information. The FRET-FLIM readout is independent of the donor or acceptor concentration making it the quantification assay of choice. Recently, FRET has been exploited to engineer sensors (FRET biosensors). FRET signal changes in these biosensors correspond to either binding or release of a ligand. How this type of readout can help uncover biological mechanisms was recently shown in a Nature Immunology article (Anzilotti 2019). The approach used in this article, and described in this webinar, opens the field to imaging in a range of situations in primary cells, where sensitivity and the need to avoid damaging the cells are both paramount.
In this tutorial on confocal imaging, you will learn how you can:
- Capture weaker signals—and still get sound, reproducible data.
- Reach faster volumetric imaging without sacrificing resolution
- Increase data throughput for your imaging needs
Learn about the new Multiplex mode for parallel pixel acquisition with the ZEISS LSM 9 family and Airyscan 2. You can now acquire up to 8 superresolution image lines with high signal-to-noise rate in a single sweep. Capture dynamic processes, cellular signaling, molecular trafficking, and diffusion events with real-time superresolution and high SNR.
The new Multiplex mode for Airyscan 2 uses smart illumination and detection schemes for parallel pixel acquisition on a confocal microscope. Scientists can now capture weaker signals, keep their context, and get statistically sound data.
Extending Airyscan imaging to larger model systems with lower expression levels, the new Multiplex mode increases acquisition speeds even further. You get superresolution and a 4 times higher SNR compared to traditional confocals. This novel concept allows rapid volumetric imaging with unprecedented resolution beyond what is available in traditional confocal systems today.
Airyscan 2 provides new data handling concepts, providing 6.6 times smaller data sizes and 5 times faster image reconstruction times. Further, optimized real time acquisition strategies employed with the LSM 9 family enable faster scan speeds for Airyscan 2, allowing higher data throughput.
In this webinar, you will learn:
- The complete workflow for in situ cryo-electron tomography
- How subtomogram averaging within the cell yields native-state structures of macromolecular complexes (e.g., the asymmetric and dilated nuclear pore of algae)
- How mapping these structures back into the native cellular environment reveals new molecular interactions that are only accessible by this technique (e.g., the binding of cargo to COPI-coated Golgi membranes and the tethering of proteasomes to the nuclear pore).
Cryo-electron tomography can visualize macromolecular structures in situ, inside the cell. Vitreous frozen cells are first thinned with a focused ion beam and then imaged in three dimensions using a transmission electron microscope. This transformative method has the power to revolutionize our understanding of cell biology, revealing native cellular architecture with molecular clarity.
In this webinar, you will learn:
- How structural biology can change the face of your research
- The biological challenges of cancer research
- Advantages of cryo-EM for your research
Covering both biology and methodology, this webinar will explain how single-particle cryo-electron microscopy enables us to gain insight into cancer development through the detailed analysis of molecular structure.
In single-particle cryo-EM, hundreds of thousands of images formed by electron scattering of individual molecules or complexes are analyzed to derive their three-dimensional structure. Technological and computational advances have dramatically transformed the field of cryo-EM in the past years, enabling structural insights at near-atomic resolution into assemblies that had not been tractable using any other structural biology technique. Consequently, cryo-EM has become a mainstream method structural biology, with a multitude of new facilities and research groups being established all over the world within just a couple of years.
This webinar will address the roles that structural biology has been playing in cancer research, uncovering cellular processes involved in cancer development and protection, and guiding drug discovery efforts. The biological challenges of cancer research will be discussed, as well as the unique strengths of cryo-EM as an experimental approach towards these questions, briefly covering the methodology and procedures in sample preparation and data processing. We will illustrate these aspects with some of the latest research from the laboratory of Eva Nogales at UC Berkeley and assess the promises and challenges of cryo-EM in our fight against cancer.
Your life sciences research often requires you to measure, quantify and understand the finest details and sub-cellular structures of your sample. You may be working with tissue, bacteria, organoids, neurons, living or fixed -cells and many different labels.
In this webinar, we will explain how Elyra 7 with Lattice SIM takes you beyond the diffraction limit of conventional microscopy to image your samples with superresolution. Learn how to examine the fastest processes in living samples – in large fields of view, in 3D, over long time periods, and with multiple colors. The new Lattice SIM technology of Elyra 7 brings structured illumination microscopy (SIM) to a new level. Groundbreaking light efficiency gives you gentle superresolution imaging with incredibly high speed – at 255 fps you will get your data faster than ever before.
See how Elyra 7 lets you combine Lattice SIM with single molecule localization microscopy (SMLM) for techniques such as PALM, dSTORM and PAINT. Choose freely among your labels when imaging with resolutions down to 20 nm laterally. High power laser lines allow you to image your sample with ease, from green to far red.
Elyra 7 is also very flexible: you can employ a wealth of contrasting techniques and combine them with optical sectioning. The new Apotome mode gives you superfast optical sectioning of your 3D samples. All that, plus Elyra 7 works seamlessly with your ZEISS SEMs in a correlative workflow.
Array tomography (AT) is a 3D image reconstruction technique for high resolution, quantitative analysis of biological structures. For optimal results, ultrathin and ordered sections are an absolute requirement.
In this webinar you will get tips and tricks to optimize the workflow of your array tomography:
- Fast and precise trimming of the sample block-face
- Adhesion of a single section to create ribbons
- Automated serial sectioning with the ARTOS 3D ultramicrotome
- Acquisition and processing of a 3D SEM dataset
- Segmentation and 3D reconstruction of cells
- Interpretation of 3D reconstructions
In this webinar you will learn:
- Improved vitrification: Specimen vitrification without synchronisation fluid
- Vitrification strategies: Optimized freezing for different techniques
- Light and electrical stimulation: Dissect cellular processes with millisecond precision
- A look into the future: freezing of crystals
Plunge freezing and cryo imaging of proteins and complexes have revealed new details in understanding the machinery of the cell and how molecules are involved in cellular processes. However, most eukaryotic cells and tissue samples cannot be plunge frozen because of the rapid decay of the cooling rate within the sample during freezing. High pressure freezing, on the other hand, is currently the main approach to vitrify larger samples (up to 200 µm) and to capture the intrinsic changes in fine structure or cellular dynamics. To further improve its cryo solutions, Leica developed a new cryo platform: the EM ICE. This new generation cryo platform combines speed, reliability and flexibility to facilitate research in various scientific fields.
The EM ICE allows users to freeze samples within milliseconds and even permits the combination of high pressure freezing with optogenetics and electrophysiology.
Join Dr. Jason Reed as he describes a novel method by which endonuclease-inhibited Cas9 can be employed as a programmable biomarker in high-speed atomic force microscopy (HS-AFM) imaging.
In this webinar, you will learn:
- How CRISPR/Cas9 can be used to ""flag"" alterations and mutations in DNA, rather than cut it
- How pairing atomic force microscopy (AFM) with optical equipment found in DVD players can be used to map DNA at a faster rate than traditional DNA sequencing
- Applications of this technology, particularly in relation to discovering and diagnosing genetic diseases
Since the diameter of the Cas9 molecule is greater than that of DNA, they are easy to locate along the DNA strand. Taking advantage of this, Jason's lab reported approximately 90% Cas9 binding accuracy to DNA molecules under optimized conditions. The alignment of single-molecule maps with nanoscale resolution becomes far more computationally straightforward than if labels are localized with multi-kb ambiguity. This process yields reduced processing time and cost for assembling a consensus map. Given its single-molecule sensitivity, approximately 15 bp accuracy, and no amplification requirement, Dr. Reed's novel method is amenable to small sample sizes. This proves to be an advantage in clinical situations where obtaining the almost 10 μg of DNA required for single-molecule sequencing is extremely difficult—if not impossible!
Join Theo Roth as he describes his lab's novel CRISPR-Cas9 genome-targeting system that does not require viral vectors to modify T cell genomes, but instead focuses on HDR. This allows rapid and efficient insertion of large DNA sequences at specific sites in the genomes of primary human T cells, and permits individual or multiplexed modification of endogenous genes. Importantly, avoiding the use of viral vectors will result in accelerated research and clinical applications, reduce experimental cost, and improve safety.
In this webinar, you will learn:
- The advantages of using HDR versus recombinant viral vectors when modifying T cell genomes
- How long double-stranded and single-stranded DNA can serve as a non-viral HDR template
- A novel method that allows for the insertion of large DNA sequences (>1Kb) without a virus!
Current efforts at reprogramming T cells for therapeutic purposes rely on using recombinant viral vectors. Unfortunately, viral vectors do not target transgenes to specific genomic sites. Moreover, the manufacturing and testing of effective viral vectors is often a lengthy and expensive process, which slows research progress and clinical use. However, recent studies have shown that re-engineering T cells in a specific and efficient manner is possible using homology-directed repair (HDR).
Join us in this webinar as Dr. Victoria Doronina helps you determine the quality of your nucleic acids. In this webinar you will learn:
- How to choose the best method to extract your nucleic acids
- Which method you should chose to determine nucleic acid quality
- How to avoid common pitfalls for both extraction and quality control
Virtually all experiments in the molecular biology lab require high quality, pure nucleic acids as a starting material. This sounds simple enough. However, as always, the trouble is in the details. There are several methods to isolate your nucleic acids. But which one is best for your experiment? How do you determine nucleic acid quality? And why does it seem to not be working?
Watch this webinar to find out the answers. Victoria will breakdown the nucleic acid purification process and how to match the right method to the right type of nucleic acid. She will also show you how to avoid common pitfalls and contaminants in isolation and quality control. This webinar is essential for anyone in a molecular biology lab who wants better results in downstream experiments!
In this webinar, you will learn:
- How to best prepare your specimen for live cell isolation by laser microdissection
- How to optimize your laser microdissection workflow
- How to avoid common pitfalls of this technique
Laser microdissection is a tool for the isolation of homogenous cell populations from their native niches in tissues to downstream molecular assays. Beside its routine use for fixed tissue sections, laser microdissection may be applied for live cell isolation. Unlike other well-established and widely used techniques for live cell isolation and single cell cloning—such as FACS, MACS, cloning by limited dilution, and so on—laser microdissection allows for capturing live cells and cell colonies without their detachment from the carrier. In other words, there is no need to prepare a single cell suspension before the isolation procedure using mechanical and enzymatic dissociation, which can affect cell fate after plating. This feature of laser microdissection is desirable for stem cell research. We established a simple strategy for the efficient live cell isolation using the Leica Laser Microdissection platform. We were able to demonstrate not only colony formation from the isolated samples containing live cells, but also single cell cloning. In this webinar, specimen preparation, laser adjustment, overall workflow, and limitations on live cell isolation by laser microdissection are discussed.
In this webinar, you will learn:
- How to overcome issues with stage drift and crosstalk between channels
- How to obtain reliable images and reproducible quantification results
- How to proceed with advanced image analysis after restoring images
During fluorescence image acquisition, many experimental uncertainties are introduced that affect the correct object interpretation and analysis. The blurring and the noise implicit in the image formation are two of the largest sources of experimental trouble. Additional aberrations, such as stage drift, and crosstalk and chromatic aberration between channels, can also affect the imaging. Huygens image deconvolution and restoration is a proven method to revert these issues and recover a more realistic representation of the original object. After restoration of the image, you can proceed with the advanced Huygens analysis options for colocalization, object measurements, and tracking.
This webinar will illustrate how you can make optimal use of the complete Huygens workflow to obtain reliable images and reproducible quantification results, focusing on the advanced analysis options offered by the Huygens software.
Join us in this webinar featuring Dr. Omonse Talton who will guide you through developing a ""fool proof"" enzyme-linked immunosorbent assay (ELISA). In this webinar, you will learn:
- When and why you should use the different types of ELISAs– direct, indirect, sandwich, competitive/inhibition ELISA
- Major considerations for developing your ELISA
- Tried and tested tips for you to perform a successful ELISA
The ELISA is one of (if not the most) common techniques in biology and biochemistry laboratories. You can use an ELISA to detect minute amounts of protein for medical diagnostics, for testing food for common allergens, and in toxicology screens screen for certain drugs. While the overall premise of an ELISA is simple, as with any assay, its success hinges on the experimental set up.
In this webinar, Omonse will give you a crash course on that experimental set up—from start to finish. She will discuss the different types of ELISAs, the major considerations in development of the assay, and troubleshooting—including tips and tricks. With this webinar, you will be able to ensure that your ELISA provides reproducible and publishable results!
In this webinar, Dr. Stephen Pettitt explains how he applies genome-wide targeted mutagenesis screens to elucidate the genetic basis of drug resistance. Using mouse and breast cancer cell lines, Dr. Pettitt’s team developed a targeted, genome-wide mutagenesis screen to identify mutations responsible for resistance to the potent PARP inhibitor talazoparib (BMN 673). The screen yielded one particularly interesting point mutation in the PARP1 gene. This mutation disrupted the ability of PARP1 to bind DNA, demonstrating that DNA binding is necessary for the action of talazoparib. Dr. Pettitt will describe how he then employed a high-density, focused sgRNA library targeting PARP1 to generate further mutants that he used to elucidate details of the structure-function relationships of PARP1. This research is not only important for unravelling the mechanisms underlying drug resistance, but it may improve future treatment plans for cancer patients.
In this webinar, you will learn:
- How to use genome-wide CRISPR screening for mutant discovery
- How to create a highly diverse, sgRNA library from Twist Bioscience for targeted, subtle mutations
- How knowledge of the structure-function relationships of PARP1 mutants can inform treatment of cancer patients with these drugs
We’ll discuss specific assays in each of these categories, the joys and pitfalls of each assay, and recommendations on how to choose the best method. You will learn tips and strategies for successful assay development using the following methods:
Proliferation:
3H-Thymidine Uptake Bromodeoxyuridine Uptake (BrdU) ATP Luminescence Fluorescent Dye Reduction (CFSE) Cytokine Measurement:
Multiplex vs. Single Cytokine Choice of Cytokine (IFNg, TNFa, IL-6, IL-1?, etc.) Kinetics of Cytokine Release Surface Antigen Expression:
CD69, CD25, PD-1, etc. Combine with CFSE, Ki67 or BrdU Kinetics are Important Cytotoxicity:
Two-Label Flow Cytometry Calcein AM Dye Release Luciferase Transduced Targets Annexin V
Activation of immune cells is the all-important first step in mounting an immune response. Immune cell activation is a popular area of research because so much happens that is key to the downstream goal of fighting infection, cancer, and disease.
There are many ways to measure immune cell activation, and they all have utility. Methods can be grouped into four main categories: Proliferation Assays, Cytokine Measurement, Surface Antigen Expression, and Cytotoxicity.
While next generation sequencing enables researchers to unveil expression levels of the entire genome, qRT-PCR remains the gold standard for measuring transcript levels of individual genes for functional studies and for the purposes of publication. In this webinar, you will learn:
• Low (1-5 genes) vs medium (~300 genes) throughput experimental design • Pros and cons of self-designed vs “off the shelf” assays • How to set up your wet lab experiments start to finish • Downloadable example step-by-step experiments with real data analysis and tutorial • Biological considerations (time series data, cell population frequency changes + more) • Examples of these techniques in publications • Common pitfalls and how to avoid them • Limitations of the technique • MIQE and publication standards
Whether you are interested in a few genes or a few hundred, join Matthew Mule as he takes you through the necessary steps to validate expression levels of target genes using qRT-PCR with single gene assays and other medium-throughput platforms.
Join us in this webinar featuring Dr. Vicki Doronina as she takes you through vital components of assay design. In this webinar you will learn:
How to choose between immortalized cells and primary cells for your assay How to avoid sources of bias in your cell-based assays How to use high throughput assays, so you can achieve greater reproducibility
By now, you have heard about the reproducibility crisis—the inability of scientists to reproduce experimental results. This crisis spurred journals to institute new requirements for publication and funding agencies to introduce more stringent rigor and reproducibility criteria. In this webinar, Vicki will address an often overlooked source of experimental variability: state of your cell lines and their external conditions.
The good news is that many of these factors can be addressed through assay design. She will show you how to move from poor reproducibility of so-called “artisan experiments” to the use of standard conditions for your cell lines and use of automatic systems for high throughput screens.
Join us in this webinar featuring Dr. Marlon Stoeckius as he explains how you can improve your single-cell RNA-sequencing (scRNA-seq) experiments.
In this tutorial, you will find: -How you can run one scRNA-seq experiment with numerous protein markers in parallel -How you can increase your recovery of single cells (up to four times!) per experiment -How you can link phenotypes to transcriptomic profiles—with higher throughput methods!
The last few years have seen the scale of single cell RNA-seq experiments increase exponentially, greatly enhancing our understanding of cell biology in development and disease. It is now feasible for researchers to characterize thousands of single cells in one experiment. However, important hallmarks of immune cell states are often not detected in scRNA-seq experiments. While lower throughput methods previously allowed researchers to link phenotypes or protein expression to transcriptomic profiles, the increase in scale of modern droplet-based methods resulted in a loss of such addressability.
Here we describe two recently developed applications that utilize antibody-conjugated oligonucleotides to enhance existing scRNA-seq platforms.
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CITE-seq, which allows measurement of a potentially unlimited number of protein markers in parallel to transcriptomes.
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Cell Hashing, which enables sample multiplexing, robust multiplet detection and super-loading of scRNA-seq platforms, allowing confident recovery of 4 times as many single cells per experiment.
Reagents for performing these assays, under the name TotalSeq™ are now available from BioLegend.
qPCR is one of the most specific and sensitive tools in molecular biology, allowing the quantification of target DNA molecules present at less than 1 in 106. Next Generation Sequencing (NGS) has similar potential. However, the presence of large amounts of non-target DNA in most clinical or environmental samples precludes easy and inexpensive analysis of rare events.
In this webinar, you will learn: 1. The epigenetic differences between microbial and human/animal genomes 2. The use of restriction endonucleases to enrich for either pathogen genomes or the human genome from complex populations. 3. How the use of enrichment and concentration can improve qPCR sensitivity 4. The use of qPCR to validate enrichment from complex samples 5. The use of NGS to validate pathogen enrichment from complex samples
Join Dr. Allyn Forsyth as he describes how successful NGS analysis of complex microbiome samples can lead to the development of non-invasive colorectal cancer screening, as well as monitoring of disease states within other cancers and Alzheimer’s Disease.
Molecular interactions are key in cellular signalling. They are often ruled or rendered by the mobility of the involved molecules. We present different tools that are able to determine such mobility and potentially extract interaction dynamics. Specifically, the direct and non-invasive observation of the interactions in the living cell is often impeded by principle limitations of conventional far-field optical microscopes, for example with respect to limited spatio-temporal resolution. We depict how novel details of molecular membrane dynamics can be obtained by using advanced microscopy approaches such as the combination of super-resolution STED microscopy with fluorescence correlation spectroscopy (STED-FCS). We highlight how STED-FCS can reveal novel aspects of membrane bioactivity such as of the existence and function of potential lipid rafts, and how the new FALCON technology eases such measurements.
Elucidating meaningful, unbiased microbial community profiles from complex microbiome samples is challenging. In this webinar, you will learn:
– the sources of bias throughout the microbiome analysis workflow – practical solutions for troubleshooting your techniques – new technologies to achieve the most representative and unbiased microbiome profiles
Join Dr. Sven Reister as he guides you through a typical workflow for analysis of microbial community profiles of complex and low biomass microbiomes, and learn how to get the most complete, unbiased microbiome profiles from even your most challenging samples.
Förster (Fluorescence) Resonance Energy Transfer (FRET) has become a powerful tool to study protein-protein interactions and signal transduction in living cells. FRET is commonly read out either by detecting the ratio of the donor and acceptor intensities (sensitized emission) or by detecting the excited state lifetime of the donor, which decreases with increasing FRET (Fluorescence Lifetime IMaging or FLIM). FLIM is robust, immune to bleaching and inherently quantitative. On confocal microscopes, FLIM is typically read out by Time-Correlated Single Photon Counting (TCSPC). This requires expensive add-on hardware and is inherently very slow, necessitating accumulation of many consecutive scans to arrive at low-noise lifetime images. For this reason fast lifetime changes, such as those encountered when reading out live-cell signaling events with FRET sensors, cannot be detected. Accumulation of images and/or very slow scanning can also cause morphological artifacts in the acquired images.
We present the Leica SP8 FALCON, a new high-end confocal instrument with built-in very fast FLIM capabilities. The instrument applies a novel technology to records photon arrival times at up to 80 MHz (160 MHz if two detectors are used) using the built-in spectral HyD detectors. To arrive at such high counts rates, the instrument uses real-time 10 GHz sampling and analysis algorithms that render it largely immune to pulse pile-up. Falcon FLIM has been fully integrated in the LAS-X software, allowing convenient acquisition of FLIM time-lapses, stitched FLIM overview images, FLIM XYZ-stacks and spectral FLIM stacks, among others, as well as combinations thereof.
We scrutinized the performance of the SP8 FALCON and applied it to record agonist-induced changes in concentration of second messengers such as cAMP and Ca2+ with sub-micrometer precision at high speed- on the order of several (512x512) frames per second (FPS). To detect cAMP by FLIM, we used our newest generation of EPAC-based dedicated FLIM sensors, which use mTurquoise2 as a donor and a tandem of two monomeric dark Venus proteins as acceptors. Reducing the image format to 128x128 pixels, we acquired good quality FLIM time-lapses at 25 FPS (83 FPS using resonant scanning), allowing detection of even very fast signaling events, including e.g. the activation of G-proteins and Ca2+ sparks. The large numbers of detected photons reduce pixel-to-pixel variability in the calculated lifetimes. Finally we demonstrate that the SP8-FALCON is ideal for fast FLIM screening applications in both fixed- and live-cell formats.
If image analysis is a place you fear to tread, or if you struggle with over complicated and time-consuming microscopy image analysis workflows, this is your opportunity to go beyond those limits.
You will learn a fast, efficient and flexible approach to 4D microscopy image analysis, which yields high quality images and results.
We'll cover:
• How to handle and process large image data quickly
• How to detect cells, nuclei, membranes and cellular structures easily with interactive image analysis including the use of virtual reality for editing
• How your research can benefit from using server environments for higher throughput and collaboration with other researchers.
Join Dr. Chris Zugates as he takes you through a typical workflow of image analysis, learn how to easily process your data and find the objects of interest quickly and interactively creating meaningful results for publication.
Chris will then show how arivis can support you through the whole workflow from efficient image acquisition up to the presentation of your work via Web or via Virtual Reality to the scientific community.
Current DNA synthesis and assembly technologies give today’s genetic engineers unprecedented freedom to control every aspect of genetic design. In this webinar on DNA assembly, you will learn:
Key concepts in DNA assembly The importance of analyzing combinatorial libraries of genetic designs for natural product biosynthesis Emerging areas of research Join Dr. Michael Smanski as he discusses his group’s work in engineering natural products, specialized metabolites that are not necessary for growth or reproduction of the organism(s) that produce them. By focusing on how combinatorial DNA assembly strategies can be used to interrogate and optimize refactored gene clusters, Dr. Smanski aims to frame an example application of DNA assembly for the production of a natural product with promising pre-clinical bioactivity.
In this webinar, you will learn about the advantages of deconvolving all your images fully automatically. Discussed in this webinar:
- How to improve the quality of your images with the new Batch Express
- How to increase resolution and signal of all your fluorescent microscopy data
- How to deconvolve your new acquired images instantly
- If you are interested in improving your fluorescent microscopy images, without manual intervention, then this is the webinar for you!
The Huygens software is considered the gold standard for deconvolution and restoration of microscopy data. Its high-quality deconvolution is now accessible with a fully automated image-processing pipeline within the new Batch Express option. You simply have to select the desired Image Feeder folder and the images in the folder will be automatically deconvolved by the Batch Express within seconds. The Batch Express supports a wide range of microscope types (widefield, confocal, spinning disk, multiphoton, STED, and a variety of SPIM/Light Sheet systems) and file formats. Compatibility with your image files is not an issue. After this webinar, your images will look better than ever before!
In this webinar, you will learn how to solve a major problem in creating expression libraries from genome sequences for downstream analyses. Specifically, you will learn:
The difference and benefits of LASSO cloning over NGS How LASSO cloning allows for the multiplex cloning of large open reading frames (ORFs) of bacterial, human, and human-microbiome genomes How this technology can impact our understanding of disease, drug discovery, precision medicine, and so much more Join Dr. Lorenzo Tosi as he describes LASSO cloning, a technique that will allow for the expression of an entire set of proteins from any organism in more rapid and cost-effective manner.
The development of tools and techniques designed to sequence and understand genomes has advanced our understanding of the world at large. However, our ability to analyze the function of expressed molecules from these genomes is lagging. Even with the advent of next generation sequencing (NGS), which has changed the landscape of genomic sequencing through increased accuracy and speed, our experiments are still limited by the length of DNA targets being read.
Utilizing a pinhole-plane imaging concept, ZEISS Airyscan allows for simultaneous improvement in resolution and signal-to-noise by capitalizing on an innovative 32-channel GaAsP photomultiplier tube (PMT) array detector. Each detection channel functions as a very small pinhole to increase resolution while the overall detector design delivers better signal-to-noise than traditional GaAsP-based confocal systems. In the past, a stack of at least five z-slices had to be deconvolved to get usable images with an optical section thinner than one Airy unit. Now, the new 2D Superresolution mode for ZEISS Airyscan delivers images with the thinnest optical section (0.2 Airy units) from a single image while maintaining the light collection efficiency of a much larger 1.25 Airy unit pinhole.
Join us in this webinar on demystifying DNA assembly. In this webinar, you will learn:
- How flanking homology DNA assembly methods work
- How to use web-based software to design experimental methods for flanking homology DNA assembly methods
- How synthetic DNA fragments fit in to the DNA assembly process
Dr. Nathan Hillson will discuss methods in flanking homology DNA assembly, including Gibson, In-Fusion, and yeast TAR assembly—amongst many other related methods. Current DNA assembly methods offer many advantages over traditional (multiple cloning site, digestion/ligation) approaches, including the ability to assemble multiple fragments at once, the lack of a necessary specific restriction enzyme, and time commitment.
One part of DNA assembly is designing these experimental methods. To that end, Dr. Hillson will provide a demonstration of how to use web-based software to automate and optimize the design of protocols for these methods.
Finally, Dr. Hillson will explain how synthetic DNA fragments fit in to the DNA assembly process and how this relates to your work.
In this webinar, you will learn about the power of 3D imaging of larger samples. In particular, you will:
Gain insight in to how to use x-ray microscopy to analyze biological samples Learn the basics about sample preparation for x-ray microscopy Understand the benefits and drawbacks to different imaging conditions X-Ray Microscopy (XRM) is a relatively new technique that combines the geometric magnification of traditional micro-CT with the optical magnification of light microscopy. Using XRM you can image the internal structure of objects with fine resolution without destroying the sample. For example, the Zeiss Versa XRM system allows an unprecedented view inside samples varying in size from the mesoscale (cm) to the microscale (µm) at consistently sub-micron image resolutions.
This webinar will focus on biological applications of X-Ray microscopy. We will cover imaging calcified structures, such as bone, to soft tissues, like the intervertebral disc. You will also learn about visualizing blood vessels using vascular tracing agents. In addition, we will cover the basics of sample preparation along with the pros and cons of different imaging conditions. Finally, we will give you a sneak view into using XRM to spatially target, in three-dimensions, tissue specific structures in a whole organism for 3D ultrastructural imaging using Focused Ion Beam – Scanning Electron Microscopy (FIB-SEM) using the ATLAS 5 Correlative Workspace.
An unlikely renaissance is occurring in biological imaging, propelled by the ongoing development of serial blockface electron microscopy imaging technologies. Imaging systems based on scanning EM instruments place large-volume automated EM serial imaging within reach of most labs. How best to take advantage of the automation and versatility? Large scale connectomics studies are stunning in their scope and detail. Smaller scale SBFI applications, however, offer academic labs and preclinical researchers opportunities to make new discoveries, measure new outcomes, and detect toxic changes in comparatively unbiased experiments. In this webinar we will discuss today’s large volume SBFI technologies, some novel applications that are in current use, and consider how the expanding size of possible specimens is promoting faster throughput imaging.
In this webinar, you will learn how to use optical imaging for functional cardiac physiological mapping of transmembrane potential, calcium handling, and metabolism.
-The main aspects covered in the webinar include: -How to design an effective optocardiography system and select optimal fluorescence probes for your research needs using open source hardware and software. -Troubleshooting tips for reducing noise and motion artifacts during data acquisition -Advanced optocardiography: multi-parametric, panoramic and transillumination imaging.
Optocardiography: Optical Imaging of Cardiac Physiology
Optical mapping is a technique that is widely used to study heart rhythm disorders, as it offers a higher spatial and temporal resolution on cardiac tissue over traditional electrical mapping. Join Dr. Igor Efimov as he takes you through the steps involved for designing a real-time physiological imaging system as well as a novel, easily-reproducible panoramic imaging system. He will explain the history of optical mapping, basic principles and techniques for obtaining high-quality signals, and useful troubleshooting advice for both the beginner and experienced researcher. The video will take a look at optocardiography of whole mammalian hearts, isolated regions of the heart, and organotypic human cardiac slices. Viewers will also discover RHYTHM, an open-source software for optical mapping data analysis that was developed by the Efimov lab. Finally, the webinar will display representative results of optocardiography studies carried out in different animal models and human heart.
In this tutorial, you will learn: How to perform isolated proteomics of microscopic regions of interest How to use this technique with FFPE tissue How proteomics can help you understand molecular mechanisms in disease
We recently developed a technique that allows localized proteomics of microscopic regions of interest, such as specific cell types or neuropathological features of disease. This technique uses laser capture microdissection (LCM) to isolate regions/cells of interest followed by label-free quantitative mass spectrometry (LC-MS). Importantly, we optimized this technique to use formalin-fixed paraffin embedded (FFPE) tissue, so that archived human tissue specimens collected at autopsy could be used. This is a particular advantage of our methodology, as the vast majority of human tissue specimens are FFPE blocks, which are currently an underutilized, but exceptionally valuable resource for medical research. We have successfully used this technique to analyze the proteome of neuropathological features that define Alzheimer’s disease (amyloid plaques and neurofibrillary tangles), as well as specific populations of neurons that are vulnerable in AD. Going forward, the use of localized proteomics has the potential to greatly increase our understanding of the molecular mechanisms that underlie AD. More broadly, this technique could be used to analyze regions or cells of interest isolated from any FFPE tissue, and therefore could be widely used to examine disease pathogenesis across a broad spectrum of diseases.
A major impediment in the determination of high resolution protein structures by single particle cryo-electron microscopy has been the presence of sample heterogeneity. Oftentimes, heterogeneity is due the dynamic nature of protein complexes, which can exist in multiple different conformational states in solution. Recent advancements in cryo-electron image processing have provided tools to accurately sort vitrified protein complexes into distinct populations that can be subsequently used to determine structures corresponding to each of the subpopulations. Such sorting algorithms have allowed high resolution structures of multiple conformations to be elucidated from a single cryo-electron microscopic grid. Besides sorting different conformations, classification can also be used to assess the influence of specific perturbations upon conformational state at level of individual protein complexes. By modulating the concentrations of different components of the system prior to vitrification, it is possible to measure the fraction of complexes belonging to each conformational state as a function of the concentration of each component and thus characterize how specific components influence the equilibrium between conformational states.
In this webinar you will learn:
- Available tools to accurately sort vitrified protein complexes into distinct populations
- How to use those classifications to assess the influence of specific perturbations upon conformational state at level of individual protein complexes.
- How to characterize how specific components that influence the equilibrium between conformational states
Recent advances in high-throughput multi-beam scanning electron microscopy (EM) and mouse whole-brain EM preparation and collection on tape (“Brain-on-Tape”) have resulted in substantial progress towards a nano-scale map of the whole mouse brain. These maps can be used to determine how individual neurons are synaptically connected and can be used to reconstruct the precise “wiring diagram” of the whole mouse brain. We discuss the methods, recent results and remaining challenges. The ZEISS MultiSEM family features 61 or even 91 electron beams scanning in parallel, resulting in unprecedented imaging speed. This finally enables extremely large-scale electron microscopy projects such as the mapping of the brain’s neural networks at high resolution. This talk will outline the operation principle of the technology and give an overview of ongoing further application developments.
In this webinar, you will learn how synthetic biology can be used to engineer desirable and robust behaviors or traits into model organisms. The main aspects to be covered in this webinar include:
Introduction to synthetic biology – what it is and what it’s not Popular applications for synthetic biology in research The pros and cons of synthetic biology Tips and resources for getting started in this field The Sky Is the Limit with Synthetic Biology! The notion of synthetic biology began to receive attention in the 1970s with the discovery of restriction endonucleases. However, the field had its real debut in 2000, when research groups in the US used synthetic biology techniques to devise biological circuit devices controlling gene expression and biological clocks by combining different genes in E. coli.
Although no standard definition exists, synthetic biology is generally understood to be a graft of engineering and biology, and this has led to some confusing and unhelpful metaphors. In this webinar, Dr. Richardson will introduce you to the interdisciplinary world of synthetic biology, busting some of the popular myths about this field—DNA is not a programming language, and cells are not compilers or circuit boards!
Dr. Richardson will also tell you about how synthetic biology differs to recombinant DNA technology, and you will get a solid overview of where and how synthetic biology can be used within research. You will receive tips and resources to help you embark on synthetic biology. The webinar will finish with open questions and future perspectives for this exciting and fast-growing field.
Expression and Purification of an Engineered, E. coli-expressed Leishmania donovani Nucleoside Hydrolase with Immunogenic Properties Potential recombinant protein vaccine candidates must meet several criteria:
They must be expressed at sufficiently high levels in the organism of choice They must be purified to high purity from the expression system in an immunogenic form They must induce potent immune responses Dr. Patrick McAtee will take you through these vaccine development steps using the nucleoside hydrolase antigen from Leishmania donovani as an example. He will demonstrate how his lab cloned and expressed the full-length, 36-dKa protein. He will discuss purification of the protein to >99% purity using anion exchange and gel filtration chromatography. He will also talk about the steps taken to ensure protein integrity and enzymatic activity using lithium dodecyl sulfate polyacrylamide gel electrophoresis (LDS-PAGE), mass spectrometry (MS), and enzymatic assays.
Dr. McAtee will then take you through in vivo testing of the vaccine candidate including analyzing antibody levels from mice immunized with the protein alone or in a stable emulsion with glucopyranosyl lipid adjuvant (GLA-SE). He will describe characterization of the type of cellular immune response induced by the protein. Finally, he will demonstrate protective efficacy in mice challenged with Leishmania mexicana.
In this webinar, you will learn how to maximize your genome editing efficiency using CRISPR/Cas9 and how to apply this technique in your research.
The main points in the webinar will include:
How to design guide RNAs using online tools specific to the genome and application of interest. Tips and practical advice to assist you in choosing and optimizing a CRISPR/Cas9 delivery system. How you can create precise mutations using homology-directed repair, including template design and cleavage site An Essential Guide to CRISPR/Cas9 Editing Efficiency While CRISPR/Cas9 editing is utilized in a wide variety of cell types, editing efficiency continues to pose a challenge to researchers. Join Dr. Allison Mayle, as she shares best practices for increasing CRISPR/Cas9 editing efficiency. Viewers will discover online tools to aid in CRISPR/Cas9 design and delivery and tips for optimizing your CRISPR/Cas9 experiments.
In this webinar, Dr. Mayle will review the factors influencing genome editing, including target sequence selection and CRISPR delivery methods. A comparison of plasmid and viral vector delivery will be provided, as well as an introduction to DNA-free CRISPR/Cas9 ribonucleoprotein reagents. Additionally, Dr. Mayle will cover best practices for CRISPR knock-in mutagenesis via homology-directed repair (HDR) and applications available from new Cas9 protein variants.
Join Dr. Karen O’Hanlon Cohrt for a practical tour of multiplex PCR technology, where you will learn the following and much more:
Principle behind multiplex PCR technology Popular applications of this technology How to set up this reaction Advantages and disadvantages Multiplex PCR Can Benefit Your Research Dr O’Hanlon Cohrt will discuss the history of multiplex PCR, how the technique works, and how to set up a reaction. She will also provide advice for optimization of primers and how to detect your targets.
Multiplex PCR technology simultaneously detects multiple nucleic acid targets in a single reaction. This method is a straightforward and efficient solution for bypassing challenges associated with limited template material. As an added bonus, this technology is cost effective because fewer reactions are needed to detect disparate targets.
Karen will show you the real beauty of multiplex PCR— how you can use very little template or sample in each experiment. She will also share the applications of this technology, which include forensic science, pathogen identification, and genetic testing—especially in cancer.
The rise of open access is changing how research is communicated. In this webinar, we’ll celebrate Open Access Week 2016 by taking a closer look at how open access affects how researchers write and publish their results. Specifically, we will:
Define open access and Creative Commons licensing and dive into the numbers about open access (journals, articles, fees) Discuss the pros and cons of open access and dispel some myths Describe how funder and government mandates will affect the open access movement and individual researchers
In this webinar, you will learn about color reproduction in microscopy images—and how to fix it if your color reproduction goes awry.
The main points we will cover are:
The reasons that could color reproduction go wrong in your microscopy images How to correct your microscopy images with inappropriate color reproduction How to be ethical with the color in your microscopy images Advice on Color in Microscopy Images Mark Clymer, an expert in microscopy imaging, will guide you through the complicated area of color reproduction in microscopy. He will offer advice and tips on how to get the most from your microscopy camera.
Color imaging is everywhere today and is often taken for granted. Images are instantly captured on cell phones, tablets, web cams, and microscopes. Pictures are shared with friends and colleagues even faster. We thrive on instant gratification. For some of us, our images are simply a snapshot in time—a selfie in front of a landmark, an amazing meal presentation, a pic showing viable cells in culture. But for others, images need to be masterpieces, expressing not just artistry but scientific discovery.
Regardless of your philosophy, we are at the mercy of the technology: cameras AND software. So as scientists, how do we ensure that we capture and communicate images that are worthy of our research? In this webinar, you’ll learn about how color in images goes awry, and what you can do about it. And there’s a twist at the end about an often overlooked villain in this whole scheme.
About the Presenter: Mark Clymer is a veteran of the microscopy industry, establishing his fundamental skills in biotech and the drug discovery labs at Sanofi, and honing those skills as product manager at Olympus, managing the core microscope product line in the Americas. Along the way, Mark attended microscopy-based courses at the Marine Biological Laboratory in Woods Hole, MA, and now shares his expertise with students during the Immunohistochemistry & Microscopy course held each March at the MBL.
Mark offers consulting services to microscopy and biomedicine-based companies, and is the former Director of Marketing for the Datacolor Scientific division of Datacolor Inc., the Swiss-headquartered market leader in color management for industry and photography. He is the author of numerous articles and blogs on microscopy and color management. You may have seen some of his posts to microscopy and imaging-based groups on LinkedIn. Mark is also an independent microscopy and spectrophotometry sales specialist for Laxco Inc.
Mark invites you to look him up on LinkedIn
Incorporating New Dyes to Simplify Panel Design: Accelerating Reagent Availability
In this webinar you will learn how custom reagents can aid you in flow cytometry panel design—without long waits for generating these components.
We will cover:
How quality custom reagents in small sizes will make your panel design easier How you can receive these reagents fast and without traditional wait times for manufacturing How you can be confident that quality control is maintained for these custom reagents Incorporating New Dyes to Simplify Panel Design Dr. Jorg Ruhrer, Director of Research Reagents at BD Bioscience will take through how to overcome a major limitation in multicolor panel design for flow cytometry experiments—limited antibody-fluorochrome combinations. Join him in this webinar as he shows you how custom reagents provide increased options in your multicolor panel design.
Flow cytometry continues to be a critical tool for the analysis, characterization, and isolation of single cells from a heterogeneous cell population. The rapid introduction of dyes based on Sirigen technology has dramatically increased the total number of parameters that can be analyzed simultaneously from a single sample. Using these dyes coupled with recent advancements in flow cytometers, it is now possible to probe more than 28 different fluorescence parameters simultaneously, allowing for a much deeper biological understanding of a sample.
However, in spite of these advancements, one of the major limitations to designing optimal multicolor panels is the unavailability of many reagents and specificities in these new fluorochromes. To address this need, BD Biosciences has launched a new product line that will give scientists immediate access to hundreds and eventually thousands of new antibody/bright dye combinations. These new combinations will give the flexibility that you need to design a multicolor panel without compromising on results.
Jorg will discuss how these quality reagents will enable easier panel design, resulting in robust high content panels that will generate quality data to drive ever deeper scientific discoveries.
In this webinar, you will learn about the process for genotyping cells—particularly tumor cells—and practical advice for analyzing the resulting data.
The main points that we will cover are:
Practical advice and methods for sample collection and preparation for your genotyping experiments An overview of the available data analysis methods for your genotyping results Examples of pathway analysis methods for determining potential mechanisms within your system Advice, tips, and applications for your genotyping experiments Dr. Ania Wronski, a breast cancer researcher from Tufts University, will guide you through the process of investigating the genotype of tumor cells. Join her in this webinar as she takes you from sample collection to data analysis.
The American Cancer Society predicts that 1.7 million new cancer cases will be diagnosed and almost 600,000 cancer deaths will occur this year in the United States alone. These staggering numbers prompted the creation of the National Cancer Moonshot initiative in the US and contributed to the prominent position of cancer research in the European Union’s Horizon 2020 project.
Recent evidence suggests that cells within a particular tumor are not all genetically identical. Most tumors exhibit some degree of cellular heterogeneity. While most cells within the tumor possess one or more dominant mutations, sometimes there is a subpopulation with different ones. Researchers hypothesize that these subpopulations might be responsible for resistance to therapeutic agents. Discovering the genotype of tumor cells might yield new therapeutic targets to aid in the fight against multidrug resistance of cancer.
Ania will use her own research as an example and give you a broad strokes overview of the steps necessary to interrogate the genotype of tumors. She will lead you through the collection and preparation of samples and discuss available data analysis methods. Finally, she will share examples of pathway analysis that can be used to discover potential mechanisms.
In this webinar you will learn the tips and tricks necessary for you to successfully work with miRNAs, and how you can use them to further your research.
The main points we will cover are:
Tips and practical advice to help you work with miRNAs, including isolation and delivery The biology and mechanism of miRNAs and why this matters to your research An example of how miRNAs are being used in disease research. The presenter will take you through the identification of certain aberrantly expressed ones in breast cancer and how they might be used as therapeutic agents. How you can identify important miRNAs in your samples—including tumor cells Practical tips, applications and advice for miRNA research Dr. Brian Adams is a researcher on the leading edge of miRNA research. Join him in this webinar as he demystifies working with miRNAs and gives you his best tips and tricks for working with them in your research.
Since Drs. Andrew Fire and Craig Mello won the Nobel Prize for their work on small RNAs, the field has exploded with possibilities and has spawned several vibrant research areas.
Brian’s own research examines the role of miRNAs in breast cancer, and the potential therapeutic opportunities they offer. He will discuss how he has identified several ones that are aberrantly expressed in triple-negative breast cancer, for which there is currently no targeted treatment options available, how reintroducing these particular ones can promote anti-tumorigenic phenotypes, and how they could be used to sensitize tumors to chemotherapeutic agents as well as gamma-irradiation.
Using this research story as an example Brian will provide you with practical, hands-on advice that will help you get better results with miRNA isolation and delivery in your experiments. He’ll also guide you through the biology and mechanisms of action of and explain the potential therapeutic applications.
Learn more about the full potential of miRNAs and find out all the inside tricks for working with them to take your research to the next level.
For more information, visit this webinar at Bitesize Bio https://bitesizebio.com/webinar/all-about-mirnas-practical-tips-advice-and-applications/
Every individual harbors millions of genetic variants, many of which may contribute to phenotype. As the cost of sequencing has plummeted, we now have the opportunity to profile this variation across thousands of individuals. Here I will provide an overview of the workflow to analyze various types of genetic variation, starting from raw sequencing data and ending with high quality genotypes. I will first present state of the art methods for mapping raw sequences to the human reference genome and calling single nucleotide polymorphisms (SNPs) across one or thousands of samples. I will then briefly describe tools for genotyping more complex variant types, focusing on STRs as a case study. Finally, I will give an overview of tools for visualizing sequencing data and for downstream analysis of genotypes.
The current state of the academic job market has come under scrutiny in recent years, largely because of the shrinking number of tenure-track jobs available. This situation has forced graduate students and postdocs to reconsider their employment options. Other scholars may realize that academia is no longer a good fit for them but are unsure of how to land a non-faculty position. This process of self-reflection and job searching can be filled with anxiety and fear.
But it doesn’t have to be.
The presenter, Heidi Scott Giusto, will share how she came to terms with something unexpected: after many years spent pursuing a PhD, she realized she didn’t want to be a professor or researcher.
Heidi will discuss how she forged her own career path outside of academia as well as 5 steps you can begin taking NOW that will help you transition when you are ready.
Next generation sequencing (NGS) is a powerful tool for investigating genome-wide phenomena. It has been used to study epigenetic profiles as part of the ENCODE project. It is being used to understand the genetic basis of both common and rare diseases. The Cancer Genome Atlas (TCGA) used NGS to map hundreds of cancer genes. There is no area of modern genetic research that has not been transformed by the advent of NGS. With continued improvements of throughput and yield, the number of human genomes that will be sequenced in the next few years is staggering.
For all the widespread uses of NGS, there are a variety of ways to end up at your destination, along with some unique challenges along the way. To determine the path, it is best to begin with the end in mind.
Whether it’s equipment costing hundreds of thousands of dollars and requiring millions of dollars in R&D investments or standardized kits that relieve you of tedious grunt work day in and day out, you rely on the support of companies like our sponsors to provide you with the equipment and supplies you need. But beyond the things they sell, these companies know a thing or two about your work-a-day world. In this webinar we will discuss how we get them to “give it up” and freely share the wisdom they have accumulated in their experience, making your work more productive, more reliable and less stressful.
All mRNA molecules recruit specific proteins to form ribonucleoprotein complexes (mRNPs). Composition and localization of many mRNPs change dynamically from translation to decay. Microscopic techniques with high spatial and temporal resolution are invaluable for studying mRNP biogenesis.
We have developed new tools based on fluorogenic forced intercalation (FIT) probes for RNA detection, quantification and interference in biological samples. The probes contain a thiazole orange (TO) dye introduced at a position normally occupied by a nucleobase. Upon binding to target nucleic acids, the TO dye increases its quantum yield and brightness substantially (greater than10 fold). These probes detect mRNA in a rapid, wash-free FISH setup using conventional wide-field microscopy. It is an ideal tool for RNA localization screens.
Nuclease resistant FIT probes containing a locked nucleic acid adjacent to the TO dye are bright and contrasted enough for use in live imaging. These probes can also be designed to target functional elements of RNAs to test the role of those in RNP biogenesis.
Absorption and emission spectra of TO are sufficiently different from EGFP to enable high sensitivity and specificity RNA-protein co-localization analysis, even with super-resolution, to study the RNA interactome. LNA modified FIT probes are excellent subjects for STED microscopy as duplex formation greatly increases their fluorescence lifetime.
We know that cellular heterogeneity in the tumor microenvironment complicates the diagnosis and treatment of cancer. Tumors are complex, dynamic systems composed of diverse cell types in various functional states, including cancer cells and infiltrating immune cells. Resolving this cellular heterogeneity requires a single-cell approach, as analyses on bulk cell preparations mask the heterogeneity of the biological system and may lead researchers down the wrong path.
Cutting-edge single-cell analysis tools for exploring the genomic, transcriptomic and proteomic states of both tumor and related immune cells are enabling researchers to understand system heterogeneity, identify cells with previously unrecognized phenotypes and elucidate important therapeutic mechanisms.
Jonathan Irish will present his work on human solid tumor cytomics, which is revealing novel melanoma and immune cell subsets. He will highlight high-content single-cell approaches developed by his lab for systems immunology and cancer biology studies of human solid tumors, including melanoma and brain cancer. Dr. Irish will also discuss technical and biological quality controls, computational analysis and the strengths of combining single-cell approaches, such as mass cytometry, phospho-flow, imaging, transcript profiling and sequencing. These cytomic approaches are especially powerful for dissecting cellular mechanisms of treatment response, monitoring key biomarkers and precise targeting of clinically relevant cell subsets.
Manisha Ray will guide you through the newest Fluidigm technologies for single-cell analysis at the genomic, proteomic and transcriptomic levels.
Biobanks, or biorepositories, store biological samples that are critical for research in the biomedical fields. Samples from biobanks have been used for genomics research, investigations into the development of personalized medicine, and treatment of diseases.
After obtaining precious specimens, it is important to not only store them for easy retrieval, but is also vital to treat the samples so there are no complications for downstream uses. Tissues, blood, and cells all have unique requirements for maintaining specimen integrity.
Leica Microsystems Laser Microdissection systems are the method of choice for laser-based microscopic sample dissection and collection. Laser Microdissection (LMD) is a microscopic technique for isolating homogeneous, specific and pure targets from heterogeneous samples for downstream analysis (DNA, RNA & proteins). In addition, the Leica Microsystems LMD systems can be used with live cell cultures (LCC, e.g. cloning) and as a manipulation tool (e.g. for live cell/organism manipulation, NanoSIMS or CLEM preparation).
Leica Microsystems recently launched new versions of their LMD systems, the Leica Microsystems LMD6 and LMD7. The new stands offer a bigger field of view at the camera ports, plus the choice between Halogen and LED transmitted light. Combined with a fresh new design, the newest Leica Microsystems LMD systems are an exciting choice for high performance and uncompromised quality.
Learn about the advantages of using Leica Microsystems LMD techniques for precise, contamination-free isolation of specific cell types. Using brain or plant tissue sections as an example, this webinar will provide an overview of the scientific and practical considerations for obtaining highly pure material for further molecular analysis in the field of Parkinson’s disease and plants.
Enjoying your work should be one of life’s top priorities not only because you spend much of your time working, but also because if you enjoy your work you’ll inevitably do a better job.
Having worked in an uptight academic lab, a very laid-back Danish biotech company, a typically British biotech lab and been my own working-from-home boss for a number of years I have tried many different approaches and structures for my day, work and life.
Although it is still a work in progress, the result is that I have some concrete guidelines that I know make my work more enjoyable and productive. In this webinar I’d like to share them with you so you can try them out to see if they work for you too.
Seeing and believing that the dress is yellow may not be that important for the casual wedding image, but in microscopy imaging seeing and believing is critical. Microscopy images that contain color, such as images from histology-stained specimens, must be compared and analyzed. If the color reproduction is poor, then there’s a chance that important information will be overlooked, not seen, and not believed.
Living cells and organisms often suffer from the high light intensities that are used in conventional imaging. Light sheet microscopy reduces phototoxic effects and bleaching, by only illuminating a specimen in a single plane at a time whilst the signal is detected in a perpendicular direction. In combination with high-speed cameras for image acquisition, light sheet microscopy is a very gentle method to observe fast biological processes in sensitive organisms over an extended time period. By moving the sample along this plane, specimens are optically sectioned and imaged in 3D. These exciting possibilities led Nature Methods to cite light sheet imaging as their Method of the Year for 2014.
In this webinar, you’ll get:
Practical advice for sample preparation, qPCR setup and result analyses Guidance on choosing the correct fluorescent labeling system for your qPCR Tips to troubleshoot the most commonly encountered issues in qPCR Join Dr. Karen O’Hanlon Cohrt for a practical tour of the process, applications and practice of qPCR, from start to finish. Beginning with an overview of how qPCR works, the webinar will give practical advice on choosing the right setup for a given experiment. You’ll also get essential advice for carrying out your experiment, including tips for sample preparation, PCR setup and analyses. Dr. O’Hanlon Cohrt will also suggest what you can do if things go wrong!
The webinar will also cover the main applications:
Identifying genes with altered expression in disease states, such as cancer Detecting disease-related gene expression and identifying microbes Viral genotyping and viral load determination Understanding biochemical and signaling pathways and other fundamental research Register now! Don’t miss this opportunity to brush up on your qPCR.
Recently, there has been a push to develop alternative methods to traditional invasive techniques, such as solid tissue biopsies, for disease diagnosis and disease progression, as well as therapeutic response. Liquid biopsy is a new, minimally invasive technology for detecting circulating biomarkers without the need for costly or invasive procedures. Liquid biopsy enables users to sensitively, specifically and rapidly analyze circulating free nucleic acids (cfDNA), circulating tumor cells (CTCs) or exosomes from a blood sample.
The Fast Module for ZEISS LSM 880 with Airyscan: Confocal Superresolution Imaging with Four Times the Speed and Improved Signal-to-Noise Ratio
First introduced in August 2014, the Airyscan detector from ZEISS represents a new detector concept for laser scanning microscopy (LSM) that enables a simultaneous resolution and signal-to-noise (SNR) increase over traditional LSM imaging. The Airyscan detector design substitutes the conventional LSM detector and pinhole scheme for an array of 32 sensitive GaAsP detector elements, arranged in a compound eye fashion that resides in the pinhole-plane while still generating an optical section. The new detection geometry allows for the collection of the spatial distribution of light originating from every point of a microscopic fluorescent object at the pinhole allowing access to higher frequency information and while additionally collecting more light for ultra-efficient imaging. Based on the Airyscan detection concept, the next innovation from ZEISS has been developed with the introduction of the Fast mode for Airyscan. The Fast mode concept utilizes the Airyscan detector technology in combination with an illumination shaping approach to enhance acquisition speeds by four times while simultaneously increasing SNR and resolution overcoming the traditional compromises of confocal imaging.
Chromatin Immunoprecipitation (ChIP) is a powerful technique for evaluating the interactions of proteins with specific regions of genomic DNA, helping to better understand the mechanisms such as, gene regulation, DNA replication, repair and recombination, and epigenetic silencing. The use of ChIP in conjunction with NGS (ChIP-Seq) has enabled wide scale application of this technique to ascertain genome wide DNA binding sites. However, sample preparation for ChIP and ChIP-seq has multiple steps which are critical to the success of the experiments and affect the reproducibility, bias, and sensitivity of the technique.
Successful ChIP experiments demand chromatin that is sheared to manageable fragments sizes while retaining the integrity of the DNA, preserving protein epitopes and the formaldehyde cross-links attaching the proteins of interest to the DNA to deliver sensitive and reproducible results. Current methods for shearing chromatin to the desired size for immunoprecipitation in ChIP experiments are a source of significant imprecision, adversely affecting the reproducibility of ChIP experiments. In this webinar, scientist from Covaris will discuss how to better optimize your sample preparation to improve the reproducibility of your results and present data demonstrating how Covaris’ AFA Technology is ideally suited to provide the highest quality sheared chromatin. Shearing chromatin with AFA preserves precious epitopes, maintains DNA quality, protects from shearing biases, and provides highly reproducible results which are the reasons AFA is quickly becoming the standard chromatin shearing solution in leading epigenetic labs.